Sertoli cells are the major component of the spermatogonial stem cell (SSC) niche; however, regulatory mechanisms in Sertoli cells that dictate SSC fate decisions remain largely unknown. Here we revealed features of the N⁶-methyladenosine (m⁶A) mRNA modification in Sertoli cells and demonstrated the functions of WTAP, the key subunit of the m⁶A methyltransferase complex in spermatogenesis. m⁶A-sequencing analysis identified 21,909 m⁶A sites from 15,365 putative m⁶A-enriched transcripts within 6,122 genes, including many Sertoli cell-specific genes. Conditional deletion of Wtap in Sertoli cells resulted in sterility and the progressive loss of the SSC population. RNA sequencing and ribosome nascent-chain complex-bound mRNA sequencing analyses suggested that alternative splicing events of transcripts encoding SSC niche factors were sharply altered and translation of these transcripts were severely dysregulated by Wtap deletion. Collectively, this study uncovers a novel regulatory mechanism of the SSC niche and provide insights into molecular interactions between stem cells and their cognate niches in mammals.

支持细胞是精原干细胞 (SSC) 生态位的主要组成部分；然而，决定 SSC 命运决定的支持细胞中的调节机制在很大程度上仍然未知。在这里，我们揭示了支持细胞中N ⁶ -甲基腺苷 (m ⁶ A) mRNA 修饰的特征，并证明了 WTAP（m ⁶ A 甲基转移酶复合物在精子发生中的关键亚基）的功能。m ⁶ A 测序分析从6,122 个基因（包括许多支持细胞特异性基因）内的15,365 个推定的富含m ⁶ A 的转录物中鉴定了 21,909 个 m ⁶ A 位点。有条件删除Wtap支持细胞中的 ssc 种群导致不育和逐渐丧失。RNA 测序和核糖体新生链复合物结合 mRNA 测序分析表明，编码 SSC 生态位因子的转录本的选择性剪接事件发生了急剧变化，并且这些转录本的翻译因Wtap缺失而严重失调。总的来说，这项研究揭示了 SSC 生态位的新调节机制，并提供了对干细胞与其在哺乳动物中的同源生态位之间的分子相互作用的见解。